Reagent Kit, Staining Method, and Application for Dual Immunohistochemistry Combined with Elastin Fiber Staining

Abstract

A reagent kit, staining method, and application for combining dual immunohistochemistry with elastin fiber staining. The reagent kit includes a combination of primary antibodies, consisting of anti-CK7 and anti-CD34 monoclonal antibodies from different species, such as rabbit anti-human CK7 monoclonal antibody and mouse anti-human CD34 monoclonal antibody. By selecting CK7 and CD34 as the primary antibody combination, the growth pattern of tumor epithelium can be accurately indicated, and the proliferation and thickening of alveolar septal fibrous tissue caused by tumors can be clearly revealed, as opposed to structural changes of alveoli induced by other factors. Combined with independent Victoria blue staining, the elastin fibers of the walls of arteries and veins, as well as the pleura of the lungs, can be clearly visualized, aiding in the assessment of the integrity of elastin fibers on the lung surface and in determining the extent of tumor infiltration.

What is claimed is: 
     
         1 . A reagent kit for dual immunohistochemistry combined with elastin fiber staining, characterized by: including a combination of primary antibodies, which consists of monoclonal antibodies against CK7 and CD34 from different species. 
     
     
         2 . The reagent kit for dual immunohistochemistry combined with elastin fiber staining according to  claim 1 , characterized by: the combination of primary antibodies includes rabbit anti-human CK7 monoclonal antibody and mouse anti-human CD34 monoclonal antibody. 
     
     
         3 . The reagent kit for dual immunohistochemistry combined with elastin fiber staining according to  claim 1 , characterized by: further comprising the following components: hydrogen peroxide blocking agent, secondary antibody reagent, DAB chromogenic solution, Fast Red series chromogenic agent, hematoxylin counterstain solution, bluing reagent, Victoria blue staining solution, 95% ethanol, and absolute ethanol. 
     
     
         4 . The reagent kit for dual immunohistochemistry combined with elastin fiber staining according to  claim 3 , characterized by: the secondary antibody reagent includes:
 1) a secondary antibody polymer reagent labeled with horseradish peroxidase from one species and a secondary antibody polymer reagent labeled with alkaline phosphatase from another species; or   2) a secondary antibody reagent composed of a mixture of horseradish peroxidase-labeled secondary antibody polymer and alkaline phosphatase-labeled secondary antibody polymer from different species.   
     
     
         5 . A staining method for dual immunohistochemistry combined with elastin fiber staining, characterized by: performing conventional slide baking, deparaffinization, rehydration, antigen heat retrieval, and blocking of endogenous peroxidase on tissue sections to be tested; incubating the sections with the combination of primary antibodies, followed by incubation with the secondary antibody reagent; performing chromogenic reaction using relevant enzyme substrates, followed by incubation with DAB chromogenic solution; after hematoxylin counterstaining and bluing, directly staining with Victoria blue; and finally performing conventional dehydration, clearing, and coverslipping. 
     
     
         6 . Use of the reagent kit according to  claim 1  in pathological tissue staining and diagnosis. 
     
     
         7 . Use of the reagent kit according to  claim 2  in pathological tissue staining and diagnosis. 
     
     
         8 . Use of the reagent kit according to  claim 3  in pathological tissue staining and diagnosis. 
     
     
         9 . Use of the reagent kit according to  claim 4  in pathological tissue staining and diagnosis. 
     
     
         10 . Use of the staining method according to  claim 5  in pathological tissue staining and diagnosis.